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TRANSFORMASI DAN KLONING PLASMID PJ804:77539 PADA E.coli TOP’10 Siu S.S Langden; Anto Budiharjo; w wijanarka; Wien Kusharyoto
Jurnal Akademika Biologi Vol. 6 No. 1 Januari 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Kloning dan transformasi vektor PJ804:77539 dilakukan dengan tujuan perbanyakan vektor pRHA pada sel bakteri E.coli TOP’10. Ekpresi vektorpRHA diharapkan dapat terjadi pada periplasma E.coli dan memberikan ekspresi berupa kemampuan resistensi terhadap Ampicillin. Ekspresi pada periplasma bertujuan untuk meminimalisir kerugian yang timbul pada sistem ekspresi di sitoplasma di antaranya tingkat ekspresi yang rendah, protein terpotong atau resiko kontaminasi. Sekresi protein rekombinan pada periplasma dapat meningkatkan aktivitas biologis serta tingkat kestabilan produk menjadi lebih tinggi. Proses isolasi protein yang diekspresikan pada periplasma  dapat dilakukan dengan perlakuan stress osmotik ringan sehingga menurunkan resiko kontaminasi protein sitoplasma. Ekspresi protein pada periplasma diarahkan oleh peptida sinyal pelB. Peptida sinyal bekerja menarik produk protein ke periplasma dengan cara berfusi dengan ujung N-terminal pada peptida yang terekspresi. Penanda selektif (selectable marker) yang terdapat pada PJ804::77539 merupakan Ampr, suatu penanda yang memampukan bakteri untuk resisten pada keberadaan antibiotik Ampicillin. Transformasi dilakukan sesuai dengan metode heat – shock dan diseleksi pada medium LB agar dan LB cair yang mengandung antibiotik Ampicillin dengan konsentrasi 100 mg/mL. Diperoleh koloni tumbuh pada medium yang mengandung Ampicillin dan dilakukan isolasi plasmid. Visualisasi hasil elektroforesis memperlihatkan adanya pita plasmid yang diisolasi dari E.coli TOP’10 pada gel elektroforesis.Kata kunci : Ampicillin, E.coli, pelB, periplasm dan pRHA
IDENTIFIKASI MOLEKULER TANAMAN PISANG RAJALAWE BERDASARKAN GEN INTERNAL TRANCRIBED SPACER (ITS) Firly Putri Fardilla; Hermin Pancasakti Kusumaningrum; w wijanarka
Jurnal Akademika Biologi Vol. 6 No. 1 Januari 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Banana is one type of horticultural commodities in a group of fruits that have a socio-economic value is high enough for the people of Indonesia. Bananas have different varieties, one banana type Rajalawe found in Central Java. Rajalawe molecular identification has not been done before. This study aims to determine the result of the identification of the molecular basis of  Rajalawe based on genes Internal transcribed spacer (ITS), in search of identity and kinship Rajalawe. The study was conducted by isolating DNA using a method Rajalawe Doyle & Doyle, followed by ITS gene amplification and sequencing analysis. The results of gene amplification ITS produce PCR product of 643 bp. The base sequence of the sequencing results are used for the construction of phylogenetic trees. Sequence similarity analysis Rajalawe show 95% homology with Musa balbisiana and alkaline difference of 1%. Phylogenetic tree analysis showed Rajalawe have a close relationship with Musa balbisiana. However, bananas Rajalawe has several different characters with Musa balbisiana with different base sequences by 5% whereas the base sequence homology between the banana Musa balbisiana and Rajalawe with 95%.Keywords: Molecular Identification, Pisang Rajalawe, Universal Primer ITS, Musa balbisiana.
PRODUKSI INULINASE DARI UMBI DAHLIA (Dahlia variabilis) OLEH Pichia manshurica DUCC Y-015 DENGAN VARIASI WAKTU INKUBASI DAN KONSENTRASI GLUKOSA SEBAGAI SUMBER KARBON TAMBAHAN Fathika Fitrania; MG. Isworo Rukmi; w wijanarka
Jurnal Akademika Biologi Vol. 7 No. 1 Januari 2018
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

The production of fructose from inulin by inulinase would only take one stage enzimatis reaction and yielding 95% fructose. Inulin obtained from the dahlia tubers and inulinase produced by Pichia manshurica DUCC Y-015. The production of inulinase  (E.C 3.2.1.7) can be influenced by the presence of glucose as  additional carbon source. The purpose of this research is to analyze influence of  variation glucose concentration and incubation time to production inulinase P. manshurica DUCC Y-015 in dahlia tubers substrate. Measurement of production enzyme covering activity inulinase and activity invertase. Both analyzed by DNS method and determined based on 1 µmole fructose resulting. The design used in this research were Randomized Factorial Block Design ( RAFBD ). Factor I in the form of glucose concentration 0% (G0); 0.25% (G1); 0.5% (G2) and factor II were incubation time 6 hours (T6), 12 hours (T12), and 18 hours (T18) with repetition 3 times. The data obtained were analyzed by ANOVA method. The result showed variation of glucose concentration and incubation time difference had no significant effect on production of inulinase from P. manshurica DUCC Y-015. The highest production of the inulinase was demonstrated by treatment G1T12 with  0,25% glucose and incubation time 12 hours with value of the activity of inulinase as much as 0,574 IU/mL. Keywords : Dahlia tubers,  Glucose, Inulinase, Incubation time, Pichia manshurica DUCC Y-015,
PENAPISAN DAN PEMANFAATAN RHIZOBAKTERI TANAMAN SORGUM (Sorghum bicolor L. Moench) SEBAGAI INOKULAN PEMACU TUMBUH TANAMAN Annisaa Widyasari; W Wijanarka; Budi Raharjo; Mamik Setyowati
Jurnal Akademika Biologi Vol. 5 No. 4 Oktober 2016
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Sorghum was a cereal crop that has many benefit such as food, feed, industrial, and bioenergy. Sorghum had a potency to be cultivated, but productivity of sorghum was still low both in quatity and quality. One way to increase production of sorghum  is using rhizobacteria as biofertilizer. The aim of this study is to get rhizobacteria that has the ability to produce IAA, solubility of phospat (P), Nitrogen (N) fixing, and analyze the effect of rhizobacteria inoculants for enhance sorgum plant growth. Isolation of rhizobacteria was done by diluting  rhizobacteria sorghum suspension from 10-1 to 10-5 and it were be platted on SEA medium. Isolates were screened by ability to produce IAA, solubility of P, and N fixing. Producing of IAA test was done by adding Salkowsky reagent on bacterial supernatant and measured absorbance at 530 nm wavelength. Solubility of P test was done by inoculating isolates in Pikovskaya media, while N fixing test was done on N fixing media (NFB). Isolates of rhizobacteria which had a potency to increase growth of plants were made inoculants to be applied in sorghum plants. The result of this study obtain 3 isolates i.e Sr 194.3; Sr 172.1; and Sr 209.1 which were considered effective for increase growth of sorghum. The conclusion  of this study isolates which showed the highest average plant height, root length, and dry weight Sr 194.3 isolate. The statistical analysis among the treatments showed that did not any significant differences on plant height, root length, and dry weight of sorghum age 28 days after farming. Keyword : Increase growth plants, Screening, Shorgum, Rhizophere.
ISOLASI BAKTERIOFAG Salmonella spp. dari BIOFILM pada SISTEM AIR MINUM ISI ULANG Rahayu Damayanti; Siti Nur Jannah; w wijanarka; Sri Hartin Rahaju
Jurnal Akademika Biologi Vol. 5 No. 2 April 2016
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

The public demands for the refill drinking water increases causing the development of refill drinking water industries. However, the problems occurred is no standard method to process drinking water with sterile and lack of government oversight. These cases give rise to sanitation which is the formation of bacteria pathogen forming biofilms in refill drinking water system. One of the bacteria pathogens is Salmonella. Salmonella in refill drinking water can cause diarrhea, because it can produce cytotoxin and enterotoxin. Bacteriophages are viruses that infect bacteria. This study aims to find natural isolates of bacteriophage from biofilm samples to infect Salmonella spp. in refill drinking water system. The isolates obtained is then characterized by biochemical test including Gram stain, a test Kligler Iron Agar (KIA) and api assay 20 E. The positive Salmonella spp. isolates are in the second dilution refill drinking water depot. The isolation of bacteriophage from biofilm is conducted with bacteriophages amplification and bacteriophage filtrate. The Infection test is performed by using Salmonella enterica, Salmonella 7A1 from Teluk Ambon and Salmonella spp. from refill drinking water depot. Platting is performed on serial dilutions of 10-2 to 10-10phage dilution. Positive result is characterized by the formation of plaque which is in source water samples, water product and drinking water depot. The number of plaques formed is calculated by Plaque Forming Units (PFU/mL) to determine quantification or calculation phages.
PRODUKSI INULINASE OLEH KHAMIR Pichia manshurica DUCC Y-015 PADA TEPUNG UMBI DAHLIA (Dahlia variabilis Willd.) DENGAN VARIASI KONSENTRASI MnSO4.H2O DAN WAKTU INKUBASI Berlian Abadianti; Agung Suprihadi; w wijanarka
Jurnal Akademika Biologi Vol. 6 No. 3 Juli 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Inulinase (E.C. 3.2.1.7) is an enzyme which able to hydrolyze an inulin into fructose and fructooligosaccharides.  The main application of inulinase enzyme in the food industries is as the ingredient in producing High Fructose Syrup (HFS). Moreover, the other important applications of inulinase enzyme are to produce ethanol, inulooligosacarida (IOS), fruktooligosacarida (FOS), pullulan, sorbitol, etc.  Pichia manshurica DUCC Y-015 is kind of yeast that is capable in producing inulinase in medium containing inulin. Optimization of inulinase enzyme production needs to be done to increase inulinase production, the way that could be conducted is by the addition of metal ion and optimization of incubation time. The purpose of this research is to investigate the effect of adding mangan ion (MnSO4.­H2O) and incubation time. In conducting this study, the researcher applies experimentally research by using Randomized Complete Block Design (RCBD) factorial pattern. The first factor is concentration of MnSO4.H2O, with concentration level 0 mM (M0), 0,1 mM (M1), 0,5 mM (M2). The second factor is the variation of incubation time, i.e. 6 hours (I6), 12 hours (I12), and 18 hours (I18) with three times repetition. The collected data were analyzed using ANOVA 5% signification (α= 0,05) and completed by the Duncan test. The result of analysis shows that variation of MnSO4.H2O concentration and incubation time does not significantly influential on inulinase activity of Pichia manshurica DUCC Y-015.Keywords: Inulinase, Inulin, Pichia manshurica DUCC Y-015, MnSO4.H2O, Incubation time.
Produksi Enzim Protease Aspergillus Flavus Pam-25 Dengan Variasi Ph Dan Waktu Inkubasi Indra Prawira; MG. Isworo Rukmi; w wijanarka
Jurnal Akademika Biologi Vol. 4 No. 2 April 2015
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Enzyme is a biokatalisator which can increase the speed of the reaction without join react. Protease enzyme is one of the enzymes that have a high economic value because its application is extensive in the field of industry. Protease is an enzyme that catalyzes the proteolytic peptide bonds in proteins termination. This research aims to determine the effect of pH and incubation time on the production of protease enzyme from A. flavus PaM-25.. This research aims to know the influence of the variation of the pH of the incubation time and against the production of the enzyme protease of A. flavus PaM-25. The research  was conducted at the laboratories of Microbiology Department of Biology, Faculty of Science and Mathematics, University of Diponegoro. Variables observed were protease activity, protein content and specific activity. Research using randomized complete block design (RAK) with two factors. The first factor is variation of pH, P1 (pH 7), P2 (pH 8), and P3 (pH 9), while the second factor is the incubation time T5 (5th day), T6 (6th day) and T7 (7th day) with repeated 3 times. Research data analyzed by Analysis of variance (ANOVA) and continued by Least Significant Difference test and Duncan Significant Difference Test at test level 5%. The results showed that the Aspergillus flavus PaM-25 has the capability of producing alkaline protease enzymes with pH range 7.0-9.0. The highest activity of proteases of   A. flavus PaM-25 retrieved on 7th day incubation time of treatment with protease activity value 1.94 U/mL, while pH treatment has no effect. The highest levels of protein found in the treatment of pH 7 and 5th days of incubation time of 1.05 mg/mL. The value of the highest purity of enzymes found in the combination of treatment pH 9 and 7th day incubation time  of 12.91 U/mg protein
Optimasi Linamarase pada Umbi Singkong (Manihot esculenta Crantz) dan Umbi Gadung (Dioscorea hipsida Dennst) dengan Variasi Suhu dan pH yang Berbeda Yoni Anggun Endah Kurniati; W Wijanarka; Endang Kusdiyantini
Jurnal Akademika Biologi Vol. 4 No. 4 Oktober 2015
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Singkong (Manihot esculenta Crantz) dan gadung (Dioscorea hipsida Dennst) merupakan bahan pangan yang memiliki kandungan karbohidrat tinggi. Singkong dan gadung sering dikonsumsi oleh masyarakat Indonesia sebagai pengganti nasi. Kedua bahan pangan ini berpotensi menyebabkan keracunan sianida apabila tidak diolah dengan baik. Umbi singkong dan gadung mengandung enzim hidrolitik yaitu linamarase (EC 3.2.1.2.1) dan substrat linamarin. Senyawa sianida dihasilkan akibat adanya hidrolisis linamarase dengan linamarin pada kondisi pH>6. Penelitian ini bertujuan untuk mengetahui kondisi optimal linamarase dari umbi singkong dan gadung. Parameter yang diukur adalah kadar sianida menggunakan spektrofotometri λ= 510 nm. Penelitian ini dilakukan secara eksperimental menggunakan Rancangan Acak Kelompok (RAK) pola faktorial. Faktor pertama optimasi linamarase meliputi suhu 350C, 400C, dan 450C, sedangkan faktor kedua meliputi pH 6.5, 7.0, dan 7.5. Hasil penelitian menunjukkan bahwa kondisi optimal linamarase singkong pada suhu 450C dan pH 7,5. Kondisi optimal linamarase gadung  pada suhu 350C dan  pH 7.5.Kata kunci : Singkong (Manihot esculenta Crantz), Gadung (Dioscorea hipsida Dennst), Linamarase, Linamarin, Sianida 
ISOLASI BAKTERIOFAG Escherichia coli DARI SISTEM DISTRIBUSI AIR MINUM ISI ULANG SEBAGAI ANTIBIOFILM Dani Sukma Saefunida; W Wijanarka; MG Isworo Rukmi; Novik Nur Hidayat
Jurnal Akademika Biologi Vol. 5 No. 2 April 2016
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

E. coli termasuk golongan bakteri koliform sebagai indikator kualitas air yang dapat membentuk biofilm pada sistem distribusi air minum isi ulang. Biofilm tersebut dapat menyebabkan kontaminasi dan penyebaran penyakit. Bakteriofag yang memiliki kemampuan dalam melisiskan inang dapat dijadikan solusi permasalahan tersebut. Tujuan penelitian adalah untuk mengisolasi bakteriofag E. coli dari sistem distribusi air minum isi ulang dan menguji aktivitas antibiofilm. Isolasi bakteriofag dilakukan dengan metode plaque assay, sedangkan uji aktivitas antibiofilm menggunakan metode microtiter plate assay. Sampel yang digunakan adalah biofilm dari pipa sumber air, tangki penyimpanan depot air minum dan produk air minum isi ulang. Hasil penelitian menunjukkan bakteriofag E. coli dapat diperoleh dari masing-masing sampel dan memiliki aktivitas antibiofilm. Kata Kunci : Antibiofilm, Bakteriofag, E. coli.
PENGARUH CaCl2.2H2O DAN WAKTU INKUBASI TERHADAP PRODUKSI INULINASE OLEH Pichia manshurica DUCC Y-015 DALAM SUBSTRAT TEPUNG UMBI DAHLIA Dahniar Saraswati; W Wijanarka; MG Isworo Rukmi
Jurnal Akademika Biologi Vol. 6 No. 3 Juli 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Fructose production from inulin by inulinase only need one reaction enzimatic step and produce 95% fructose. Inulin obtained from dahlia tuber and inulinase produced by Pichia manshurica DUCC Y-015. Inulinase production (E.C. 3.2.1.7.) can be influenced by metal salt suplementation, such as CaCl2.2H2O. The purpose of this research were to known the influence of CaCl2.2H2O and incubation time to inulinase production by P. manshurica DUCC Y-015 on Dahlia Tuber Substrate. The design that use in this research were Randomized Factorial Block Design ( RAFBD ). Factor I (CO, C­1, C2, C3) as the concentration of CaCl2.2H2O (0 mM, 0.25 mM, 0.50 mM, 1.00 mM) and Factor II ( T12, T18, dan T24 ) as incubation time ( 6, 12, 18 hour), the repetition were 3 times. The result analyze by ANOVA (Analysis of Variance) and continued by LSD test. The result of this research indicate that CaCl2.2H2O and incubation time were not significantly influence to inulinase production. The highest inulinase production by Pichia manshurica DUCC Y-015 indicate by C2T12 treatment which use 0.50 mM CaCl2.2H2O and 12 hour incubation time, the enzyme activity is 0.60 IU/mLKey Words : CaCl2.2H2O, Dahlia tuber,  Incubation time, Inulin, Inulinase, Pichia manshurica DUCC Y-015.
Co-Authors Adzar Rofiqoh Agung Suprihadi Annisaa Widyasari Anto Budiharjo Berlian Abadianti Budi Raharjo Dahniar Saraswati Dani Sukma Saefunida Endang Kusdiyantini Fathika Fitrania Firly Putri Fardilla Gabriela Christy Sabbathini Hermin Pancasakti Kusumaningrum Indra Prawira Khrisna Lazuardi Budi Mamik Setyowati MG Isworo Rukmi Nopvita Windi Astuti Novik Nur Hidayat Prawatya Cahyani Rahayu Damayanti Riza Laksita Devi Mutiaratri Siti Nur Jannah Siu S.S Langden Sri Hartin Rahaju Sri Pujiyanto Susiana Purwantisari Wien Kusharyoto Yoni Anggun Endah Kurniati